Surface-Functionalized Microgels as Artificial Antigen-Presenting Cells to Regulate Expansion of T Cells.

Artificial antigen-presenting cells (aAPCs) are currently used to manufacture T cells for adoptive therapy in cancer treatment, but a readily tunable and modular system can enable both rapid T cell expansion and control over T cell phenotype. Here, it is shown that microgels with tailored surface biochemical properties can serve as aAPCs to mediate T cell activation and expansion. Surface functionalization of microgels is achieved via layer-by-layer coating using oppositely charged polymers, forming a thin but dense polymer layer on the surface. This facile and versatile approach is compatible with a variety of coating polymers and allows efficient and flexible surface-specific conjugation of defined peptides or proteins. The authors demonstrate that tethering appropriate stimulatory ligands on the microgel surface efficiently activates T cells for polyclonal and antigen-specific expansion. The expansion, phenotype, and functional outcome of primary mouse and human T cells can be regulated by modulating the concentration, ratio, and distribution of stimulatory ligands presented on microgel surfaces as well as the stiffness and viscoelasticity of the microgels.

The Prognostic Effect of Immune Cell Infiltration Depends on Molecular Subtype in Endometrioid Ovarian Carcinomas.

PURPOSE: Endometrioid ovarian carcinoma (ENOC) is the second most-common type of ovarian carcinoma, comprising 10%-20% of cases. Recently, the study of ENOC has benefitted from comparisons to endometrial carcinomas including defining ENOC with four prognostic molecular subtypes. Each subtype suggests differential mechanisms of progression, although tumor-initiating events remain elusive. There is evidence that the ovarian microenvironment may be critical to early lesion establishment and progression. However, while immune infiltrates have been well studied in high-grade serous ovarian carcinoma, studies in ENOC are limited. EXPERIMENTAL DESIGN: We report on 210 ENOC, with clinical follow-up and molecular subtype annotation. Using multiplex IHC and immunofluorescence, we examine the prevalence of T-cell lineage, B-cell lineage, macrophages, and populations with programmed cell death protein 1 or programmed death-ligand 1 across subtypes of ENOC. RESULTS: Immune cell infiltrates in tumor epithelium and stroma showed higher densities in ENOC subtypes with known high mutation burden (POLEmut and MMRd). While molecular subtypes were prognostically significant, immune infiltrates were not (overall survival P > 0.2). Analysis by molecular subtype revealed that immune cell density was prognostically significant in only the no specific molecular profile (NSMP) subtype, where immune infiltrates lacking B cells (TILB minus) had inferior outcome (disease-specific survival: HR, 4.0; 95% confidence interval, 1.1-14.7; P < 0.05). Similar to endometrial carcinomas, molecular subtype stratification was generally superior to immune response in predicting outcomes. CONCLUSIONS: Subtype stratification is critical for better understanding of ENOC, in particular the distribution and prognostic significance of immune cell infiltrates. The role of B cells in the immune response within NSMP tumors warrants further study.

The magnetic field strength and the force distance dependency of the magnetically controlled growing rods used for early onset scoliosis.

Magnetically controlled growing rods (MCGR's) have revolutionized the treatment of early-onset scoliosis (EOS) because painless lengthenings can be done in the outpatient clinic without anesthesia. Untreated EOS leads to respiratory insufficiency and reduced life expectancy. However, MCGR's have inherent complications like non-functioning of the lengthening mechanism. We quantify an important failure mechanism and give advice on how to avoid this complication. The magnetic field strength was measured on new/explanted rods at different distances between the external remote controller and the MCGR and likewise in patients before/after distractions. The magnetic field strength of the internal actuator decayed fast with increasing distances and plateaued at 25-30 mm approximating zero. Two new and 12 explanted MCGRs was used for the lab measurements of the elicited force using a forcemeter. At a distance of 25 mm, the force was reduced to approximately 40% (ca. 100 N) compared to zero distance (ca. 250 N), most so for explanted rods. This is used to point out the importance of minimizing the implantation depth to ensure proper functionality of the rod lengthening in clinical use for EOS patients. A distance of 25 mm from skin to MCGR should be considered a relative contraindication to clinical use in EOS patients.

Inter- and intrarater reliability of measuring lengthening of magnetically controlled growing rods on digital radiographs.

PURPOSE: The aim of this study was to determine inter- and intraobserver reliability of delta rod extension, and total rod length measured on digital x-rays in patients with early onset scoliosis (EOS) treated with magnetically controlled growing rods (MCGR). For the last decade, patients with EOS have been treated with MCGR. Replacement of MCGR relies heavily on the measurement done at every lengthening session. Only a few studies have looked at inter- and intraobserver reliability of rod lengthening, and none have used the delta extension before. METHODS: 202 radiographs presented in random order were rated and measured twice with at least a 14-day interval and differing order of the radiographs. The measuring was done at both rods. All x-rays came from 15 patients diagnosed with EOS and treated with MCGR from 2009 until 2019. The total extension length and the delta extension (the difference in total extension length between two lengthening in succession) were measured, and the intraclass correlation coefficient (ICC) calculated for both measurements RESULTS: Intrarater ICC scores varied from moderate to good, but non-significantly. Interrater reliability increased significantly from moderate (ICC 0.72 [0.68; 0.76] and 0.73 [0.69; 0.77] to excellent (ICC 0.91 [0.88; 0.93] and 0.97 [0.96: 0.98]), when examining delta extension every sixth instead of every second month. CONCLUSION: Measuring rod lengthening on x-rays can be done every 6 months, with an ample reliability. The ICC's for the delta extension with 2-3 months interval were only moderately precise, compared to the near perfect ICC's for the total extension length.

Pro-cachectic factors link experimental and human chronic kidney disease to skeletal muscle wasting programs.

Skeletal muscle wasting is commonly associated with chronic kidney disease (CKD), resulting in increased morbidity and mortality. However, the link between kidney and muscle function remains poorly understood. Here, we took a complementary interorgan approach to investigate skeletal muscle wasting in CKD. We identified increased production and elevated blood levels of soluble pro-cachectic factors, including activin A, directly linking experimental and human CKD to skeletal muscle wasting programs. Single-cell sequencing data identified the expression of activin A in specific kidney cell populations of fibroblasts and cells of the juxtaglomerular apparatus. We propose that persistent and increased kidney production of pro-cachectic factors, combined with a lack of kidney clearance, facilitates a vicious kidney/muscle signaling cycle, leading to exacerbated blood accumulation and, thereby, skeletal muscle wasting. Systemic pharmacological blockade of activin A using soluble activin receptor type IIB ligand trap as well as muscle-specific adeno-associated virus-mediated downregulation of its receptor ACVR2A/B prevented muscle wasting in different mouse models of experimental CKD, suggesting that activin A is a key factor in CKD-induced cachexia. In summary, we uncovered a crosstalk between kidney and muscle and propose modulation of activin signaling as a potential therapeutic strategy for skeletal muscle wasting in CKD.

Magnetically controlled growing rods in early-onset scoliosis.

INTRODUCTION: Early-onset scoliosis (EOS) may result in disability and a reduced life expectancy. The aim of this study was to report the results of primary magnetically controlled growing rods (MCGR) in a consecutive group of patients with EOS diagnosed and operated at Aalborg University Hospital, Denmark, from 2009 and onwards and with at least two years of follow-up. METHODS: Data were extracted from the electronic patient records and the Picture Archiving and Communication System. All data were extracted by an unbiased observer. Demographics, any complication and the Cobb angles and maximal kyphosis angles preoperatively and post-operatively were recorded. Likewise, the total expansion of the MCGR and the increase in T1-T12 and T1-S1 heights were recorded. RESULTS: A total of 15 patients (three females) were followed for an average of 3.75 years. The Cobb angles were corrected on average by 68% and the maximal kyphosis angle by 45%. The thoracic height increased significantly with only two patients (still undergoing expansions) with a T1-S1 height below 22 cm. Four complications were recorded (one deep infection and three non-functioning rods), all resulting in rod exchange. The complication rate was 27% or 0.07 per patient per year. CONCLUSIONS: The MCGR may reduce the deformity and support thoracic and pulmonary growth without any need for repeated surgeries. The number of complications in the present series was low compared with the literature with an average of 0.07 complications per year per patient or a total complication rate of 27%. FUNDING: none. TRIAL REGISTRATION: not relevant.

P2Y2R Signaling Is Involved in the Onset of Glomerulonephritis.

Endogenously released adenosine-5'-triphosphate (ATP) is a key regulator of physiological function and inflammatory responses in the kidney. Genetic or pharmacological inhibition of purinergic receptors has been linked to attenuation of inflammatory disorders and hence constitutes promising new avenues for halting and reverting inflammatory renal diseases. However, the involvement of purinergic receptors in glomerulonephritis (GN) has only been incompletely mapped. Here, we demonstrate that induction of GN in an experimental antibody-mediated GN model results in a significant increase of urinary ATP-levels and an upregulation of P2Y2R expression in resident kidney cells as well as infiltrating leukocytes pointing toward a possible role of the ATP/P2Y2R-axis in glomerular disease initiation. In agreement, decreasing extracellular ATP-levels or inhibition of P2R during induction of antibody-mediated GN leads to a reduction in all cardinal features of GN such as proteinuria, glomerulosclerosis, and renal failure. The specific involvement of P2Y2R could be further substantiated by demonstrating the protective effect of the lack of P2Y2R in antibody-mediated GN. To systematically differentiate between the function of P2Y2R on resident renal cells versus infiltrating leukocytes, we performed bone marrow-chimera experiments revealing that P2Y2R on hematopoietic cells is the main driver of the ATP/P2Y2R-mediated disease progression in antibody-mediated GN. Thus, these data unravel an important pro-inflammatory role for P2Y2R in the pathogenesis of GN.